PUBLICATION

See-Thru-Gonad zebrafish line: developmental and functional validation

Authors
Presslauer, C., Bizuayehu, T.T., Razmi, K., Fernandes, J.M., Babiak, I.
ID
ZDB-PUB-160923-9
Date
2016
Source
Reproduction (Cambridge, England)   152: 507-17 (Journal)
Registered Authors
Babiak, Igor, Fernandes, Jorge
Keywords
none
MeSH Terms
  • Animals
  • Cells, Cultured
  • Embryo, Nonmammalian/cytology*
  • Embryo, Nonmammalian/metabolism
  • Fluorescent Dyes/metabolism*
  • Gene Expression Regulation, Developmental
  • Germ Cells/cytology*
  • Germ Cells/metabolism
  • Gonads/growth & development*
  • Gonads/metabolism*
  • Life Cycle Stages
  • MicroRNAs/genetics*
  • Zebrafish/physiology*
PubMed
27655215 Full text @ Reproduction
Abstract
Zebrafish are an important model species in developmental biology. However, their potential in reproductive biology research has yet to be realized. In this study, we established See-Thru-Gonad zebrafish, a transparent line with fluorescently labeled germ cells visible throughout the life cycle, validated its gonadal development features, and demonstrated its applicability by performing a targeted gene knockdown experiment using vivo-morpholinos (VMOs). To establish the line, we crossed the zf45Tg and mitfa(w2/w2); mpv17(b18/b18) zebrafish lines. We documented the in vivo visibility of the germline-specific fluorescent signal throughout development, from gametes through embryonic and juvenile stages up to sexual maturity, and validated gonadal development with histology. We performed targeted gene knockdown of the microRNA (miRNA) miR-92a-3p through injection of VMOs directly to maturing ovaries. After the treatment, zebrafish were bred naturally. Embryos from miR-92a-3p knockdown ovaries had a significant reduction in relative miR-92a-3p expression and a higher percentage of developmental arrest at the 1-cell stage as compared with 5-base mismatch-treated controls. The experiment demonstrates that See-Thru-Gonad line can be successfully used for vertical transmission of the effects of targeted gene knockdown in ovaries into their offspring.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping